Lab for highly sensitive blood-PCR ?

[Markus83]

I am looking for a laboratory that can perform highly sensitive PCRs from blood for various pathogens. I am interested in Chlamydia pneumoniae and Parvo B19 because they are serologically conspicuous in my case. I couldn't find them here in Germany. There are some papers that found Parvo B19 as a cause for CFS in some patients, but the concentrations were extremely low. In Germany there is no laboratory that can measure such low concentrations, not even the Charite. Hence my question, if somebody somewhere knows a laboratory, where I could send in from Germany.

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[Learner1]

I am looking for a laboratory that can perform highly sensitive PCRs from blood for various pathogens. I am interested in Chlamydia pneumoniae and Parvo B19 because they are serologically conspicuous in my case. I couldn't find them here in Germany. There are some papers that found Parvo B19 as a cause for CFS in some patients, but the concentrations were extremely low. In Germany there is no laboratory that can measure such low concentrations, not even the Charite. Hence my question, if somebody somewhere knows a laboratory, where I could send in from Germany.

Translated with www.DeepL.com/Translator (free version)

Have you talked to the folks at:

IMD Institut für Medizinische Diagnostik Berlin-Potsdam GbR
IMD Berlin MVZ

They have an extremely impressive test request sheet called specialdiagnostics.

Even if they can't do it, they may know someone who can.

However, I'm seeing one of the top US ME/CFS specialists who was an AIDS doctor for many years and very clever at testing for infections, and he told me there is no real good chlamydia pneumonia PCR test.

My antibodies looked suspicious enough, along with mycoplasma, but he prescribed 4 months of IV acyclovir, rifampin, and doxycycline 3 days a week, which he devised after talking to four other experts about my case, which dramatically helped. I had been on oral azithromycin and rifampin previously, but they weren't enough to kill them.

As for parvovirus, I've only had the antibody testing as well. But looks like Mayo offers PCR:

https://www.mayocliniclabs.com/test-catalog/Clinical+and+Interpretive/83151

 

[Markus83]

I talked to different specialized university labs and they can't do it sensitive enough for my purpose. IMD not, too (they cannot even do C. pneumoniae PCR from blood at all). Maybe it's even impossible except for research purposes.

So I need information from the labs which is the smallest amount of bacteria (C. pneumoniae) or virus (Parvo B19) copies per ml blood they can detect. For example, according to the research papers, for Parvo B19 I need a PCR which can detect down to 10 cop/ml blood, which is extremely few. Even Charité cannot do this (I had the opportunity to talk to one of the directors of the institute for viral diagnostics). But maybe it's possible in other countries.

I know that Prof. Stratton (Vanderbilt university) developed his own sensitive PCR test for C. pneumoniae, because all other tests don't work well. So that would fit what you're CFS specialist (Dr Kaufman?) said. Unfortunately it's not possible to get tested with the Stratton-PCR as far as I know.

I did test chlamydia and parvo in Germany at a university lab in Germany, but both tests came back negative. However, I'm not sure if the PCR was sensitive enough.

 

[Cipher]

I am interested in Chlamydia pneumoniae and Parvo B19 because they are serologically conspicuous in my case

What was the antibody titers for IgG/IgM?

IMD not, too (they cannot even do C. pneumoniae PCR from blood at all).

They seem to perform lymphocyte transformation test for Chlamydia pneumonia. This is a very interesting method to detect chronic infections, you can read more about it here.

 

[Markus83]

What was the antibody titers for IgG/IgM?

NS1 band positive in Western Blot for Parvo B19, which is associated in different studies with CFS. Look at some older posts of myself for more information.

Chlamydia pn: High IgG and IgA (IgA likely an indicator for chronic infection).

They seem to perform lymphocyte transformation test for Chlamydia pneumonia.

I know this test, but I do not believe that this test can reliably distinguish between active and past infection. It is claimed sometimes, but has not been proven. Seems that outside Germany this test is hardly known. So you can trust the lab (which makes a lot of money with the test), but I prefer scientific studies which are not here to date. In my eyes a positive LTT does only say that you have had contact with the pathogen in the past.

 

[Cipher]

NS1 band positive in Western Blot for Parvo B19, which is associated in different studies with CFS. Look at some older posts of myself for more information.

Old post:

Parvo came back IgM negative and IgG positve (Elisa 7.9, normal < 0.9). Wester blot IgG bands found: VP-2p, VP-N, VP-1S, VP-2r, NS1. The lab states: "NS1 band positive. Chronic persistent or past infection. To rule out chronic persistend infection PCR of blood serum recommended."

I don't know what to do with that either. 2015 I had also very high IgG-titer against parvo in another lab (144 U, normal < 3 U).

The positive IgG NS1 is very interesting, it has indeed have been linked to CFS (41.5 % of CFS patients positive vs 7% of controls). What's the name of the lab that performed the western blot? It's also interesting that the ELISA was positive but not by much (9 times the negative), while the the other lab found very high levels (48 times the negative) (144 U, normal < 3 U). Do you know the name of the laboratory that performed the very positive test and the method they used?

Old post:

Given that my IgG for Chlamydia pneumoniae was 450 (normal < 22) and IgA was 50 (normal < 22) together with my borderline lyme serology, I think I would stay on antibiotics and maybe try to add Azithromycin also.

Do you know which method was used to analyze the C. pneumoniae antibodies and the name of the laboratory?

I know this test, but I do not believe that this test can reliably distinguish between active and past infection. It is claimed sometimes, but has not been proven. Seems that outside Germany this test is hardly known. So you can trust the lab (which makes a lot of money with the test), but I prefer scientific studies which are not here to date. In my eyes a positive LTT does only say that you have had contact with the pathogen in the past.

Well, IMD Berlin have in collaboration with Charité University Medicine Berlin conducted a scientific study supporting the notion that a positive LTT strongly indicates an active infection in the context of Borrelia:

The Lymphocyte Transformation Test for Borrelia Detects Active Lyme Borreliosis and Verifies Effective Antibiotic Treatment (2012)

Abstract
Borrelia-specific antibodies are not detectable until several weeks after infection and even if they are present, they are no proof of an active infection. Since the sensitivity of culture and PCR for the diagnosis or exclusion of borreliosis is too low, a method is required that detects an active Borrelia infection as early as possible. For this purpose, a lymphocyte transformation test (LTT) using lysate antigens of Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii and recombinant OspC was developed and validated through investigations of seronegative and seropositive healthy individuals as well as of seropositive patients with clinically manifested borreliosis. The sensitivity of the LTT in clinical borreliosis before antibiotic treatment was determined as 89,4% while the specificity was 98,7%. In 1480 patients with clinically suspected borreliosis, results from serology and LTT were comparable in 79.8% of cases. 18% were serologically positive and LTT-negative. These were mainly patients with borreliosis after antibiotic therapy. 2.2% showed a negative serology and a positive LTT result. Half of them had an early erythema migrans. Following antibiotic treatment, the LTT became negative or borderline in patients with early manifestations of borreliosis, whereas in patients with late symptoms, it showed a regression while still remaining positive. Therefore, we propose the follow-up monitoring of dis-seminated Borrelia infections as the main indication for the Borrelia-LTT.

In patients with early manifestations of Borrelia who had SI>5 (SI <2 =negative; SI >2<3 = borderline, SI >3<5 = weak positive; SI>5 = positive), 78 % became negative after antibiotic treatment, while only 1.5 % remained >5. In the group of LTT-positive patients with late manifestation of Borrelia, 22 % became negative after antibiotic treatment, but only 11% remained >5. This is in line with the notion that Borrelia can persist in the human body not only in the spirochetal but also in the antibiotic-resistant biofilm form, especially if not treated early.

 

[Markus83]

Why is it important to know which labs did the testing? I don't want to write it openly because some people would (maybe) be able to trace back who I am in real life. All antibody tests were Elisas (although different labs and manufacturers) except the Western Blot.

I know the LTT study from IMD, but this was just for Lyme disease and they did not include a sufficient amount of patients with autoimmune or other infectious diseases as a control group. So you do not know much about the specifity of the borrelia-LTT test except that it is (mostly) negative in healthy controls.

 

[Cipher]

Why is it important to know which labs did the testing? I don't want to write it openly because some people would (maybe) be able to trace back who I am in real life. All antibody tests were Elisas (although different labs and manufacturers) except the Western Blot.

Well I'm interested to know from a personal level as I've not been tested for these pathogens yet and it would be helpful to know what labs perform sensitive serology tests. Maybe you can PM me the lab names?

I know the LTT study from IMD, but this was just for Lyme disease and they did not include a sufficient amount of patients with autoimmune or other infectious diseases as a control group. So you do not know much about the specifity of the borrelia-LTT test except that it is (mostly) negative in healthy controls.

But you said that you believe that LTT can't reliably distinguish between an active infection and an old infection. What did you base that on? This study contradicts that, as the levels dropped when the infection was treated.

It seems like the risk of false positives are low, even in the context of other infections or autoimmune diseases:

Therefore, 16 (1.1%) of the 1480 Borrelia-LTT results must be regarded as false positives.

In 160 seronegative subjects/patients (120 healthy seronegative and 40 patients with autoimmune diseases), a specificity of 98,7% was found if the borderline results of SI > 2 < 3 were evaluated as negative. Weak false positive reactions (SI> 3 <5) were detected in only 2 cases. No differences were found between clinically healthy subjects and patients with autoimmune diseases.

For instance, it was only possible in individual cases to examine patients with active syphilis (n = 3) or leptospirosis infection (n = 2) for potential cross reactivity. In these few cases, there was no evidence of such cross-reactivity in the Borrelia-LTT. Allergies, autoimmune diseases and acute, persistent and latent viral infections (including HIV, EBV, CMV, VZV) have now been excluded, by further investigations, as a possible cause of false-positive reactions (unpublished data).